LOCUS       sequence-436073-        7113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The base plasmid of TUNEYALI for TF47.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7113)
  TITLE     Regulating transcription factors in Yarrowia lipolytica
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7113)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCfb13237
FEATURES             Location/Qualifiers
     source          1..7113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1003..2031
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1003..2031
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     primer_bind     complement(2048..2066)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     misc_RNA        3718..3792
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(4197..4230)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(4437..4456)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4462..4481)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4523..4614
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4615..4683,4684..5475)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4833..4852)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5597..6185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6086..6105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6339..6356
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6454..6471
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6455..6471
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6941..6974
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 aaggagtttg gcgcccgttt tttcgagccc cacacgtttc ggtgagtatg agcggcggca
       61 gattcgagcg tttccggttt ccgcggctgg acgagagccc atgatggggg ctcccaccac
      121 cagcaatcag ggccctgatt acacacccac ctgtaatgtc atgctgttca tcgtggttaa
      181 tgctgctgtg tgctgtgtgt gtgtgttgtt tggcgctcat tgttgcgtta tgcagcgtac
      241 accacaatat tggaagctta ttagcctttc tattttttcg tttgcaaggc ttaacaacat
      301 tgctgtggag agggatgggg atatggaggc cgctggaggg agtcggagag gcgttttgga
      361 gcggcttggc ctggcgccca gctcgcgaaa cgcacctagg accctttggc acgccgaaat
      421 gtgccacttt tcagtctagt aacgccttac ctacgtcatt ccatgcatgc atgtttgcgc
      481 cttttttccc ttgcccttga tcgccacaca gtacagtgca ctgtacagtg gaggttttgg
      541 gggggtctta gatgggagct aaaagcggcc tagcggtaca ctagtgggat tgtatggagt
      601 ggcatggagc ctaggtggag cctgacagga cgcacgaccg gctagcccgt gacagacgat
      661 gggtggctcc tgttgtccac cgcgtacaaa tgtttgggcc aaagtcttgt cagccttgct
      721 tgcgaaccta attcccaatt ttgtcacttc gcacccccat tgatcgagcc ctaacccctg
      781 cccatcaggc aatccaatta agctcgcatt gtctgccttg tttagtttgg ctcctgcccg
      841 tttcggcgtc cacttgcaca aacacaaaca agcattatat ataaggctcg tctctccctc
      901 ccaaccacac tcactttttt gcccgtcttc ccttgctaac acaaaagtca agaacacaaa
      961 caaccacccc aaccccctta cacacaagac atatctacag caatgggtaa aaagcctgaa
     1021 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg
     1081 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga
     1141 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg
     1201 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag
     1261 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa
     1321 accgaactgc ccgctgttct gcagccggtc gcggaggcaa tggatgccat tgctgcggcc
     1381 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
     1441 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
     1501 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
     1561 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg
     1621 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
     1681 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
     1741 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat
     1801 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
     1861 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg
     1921 cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc
     1981 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata atcatgtaat
     2041 tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag
     2101 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     2161 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     2221 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgagcagg cttggaggcg
     2281 acgtggcagt ggctcgcggg aggatcggcg ggaaactccg ggatatccgt cgagagttta
     2341 cacgtgaatg ggcagcgcaa tccgttgacg acgatacgac tggcaaagta gcgacgatac
     2401 ctgccagaca ggtgacatgt gcaggccgca ctaacaagga aacgggcgct ggggggggcg
     2461 ggcttctaga ctttgccctt gaacaggaat ctagtggggg cttgtctttc cgccaatggg
     2521 ggagcgcctg ttgagcgacc gtgcatgctg gaacgccaag tgtatgtaca gctggtggtc
     2581 tcgcagcggt atgtgacggg acttacatct ctcgtttttt catgaccacg ttttcacagg
     2641 ctcggaggta cgttaaagtt ttgaaggctg catctgaacc gaggtatggg ggagtttgaa
     2701 cagcaacagt gttggggctg agggggccaa gatcggggca agcagaggtc ttagatcaat
     2761 tgtggggatc ccaaagggct cgttatcacc tttttccacc caattcgggt cccaattgat
     2821 ccactactgg cttgcccaag ttaccccaga aatgccgccc cggatttctc caaaaaccta
     2881 ataagcttca tggaacttgg tggaagtgac tttctacaga gtggagagaa ccgtggacac
     2941 gtggcaatgg cgctgaccgt gtccccgagc cgaatcgacg tgaggggaga acggagtatc
     3001 tgcggtcatg tgaccttcca gagcggcgtc gccagtgtgc acgcggtgac ccccagtttg
     3061 ggtctctgtc acacgcatac tacctcggct ctccacatgc tgaactttat ctttcgtggg
     3121 gatcataccg aaagttgcaa ctaccaggtg tatataaagc ctggtagact ccccccactt
     3181 tggacctcat ccaaccaaga cacacaaaaa tggaccgact taacaacctc gccacccagc
     3241 tcgagcagaa ccccgccaat tcgtagataa tggaatacaa atggatatcc agagtataca
     3301 catggatagt atacactgac acgacaattc tgtatctctt tatgttaact actgtgaggc
     3361 attaaataga gcttgatata taaaatgtta catttcacag tctgaacttt tgcagattac
     3421 ctaatttggt aagatattaa ttatgaactg aaagttgatg gcatccctaa atttgatgca
     3481 acggaatgcg tgcgatagtg aatcattgct aacagatcaa gtaatcgtat caatcgttcc
     3541 acgtcttatc cagactcgat ttccgcccta acctaaggaa actgctgagt aatttcgagg
     3601 ttggttccta acaaaagcca agaaacgcgc cgatggttta gtggtaaaat ccatcgttgc
     3661 catcgatggg cccccggttc gattccgggt cggcgcattg caattacata tataccggtt
     3721 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     3781 accgagtcgg tggtgctttt tttgtgtggg tacggtgcgt tggagaccca tagtcctaga
     3841 ggtttacccg agcctgggca ataattgttg ctgtcagaaa ccccagcctc aaatcctacc
     3901 ccactatagc taattttcgg tggtaattaa actgaggtca tgagttgact tatccagtgt
     3961 gtcttttccg tctttatgga ctgaagagct cttcaatgac cacctacgat ctcaactacg
     4021 actactcctt tgagtctctg gactcgtttg actacttgtc cgagagccag gtgtcgaccg
     4081 ccgccactag tcccggtgtt tatgactatt cgggctttta tggagatcac aaggcccatg
     4141 ctggaggtca cagcggcatc gcgtgcattc gcggccgcat ttaaatcccc acttcagaag
     4201 ttcctatact ttctagagaa taggaacttc actatagagt cgaataaggg cgacacccca
     4261 taattagccc gggcgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
     4321 ggcagttccc tactctcgca tggggagtcc ccacactacc atcggcgcta cggcgtttca
     4381 cttctgagtt cggcatgggg tcaggtggga ccaccgcgct actgccgcca ggcaaacaag
     4441 gggtgttatg agccatattc aggtataaat gggctcgcga taatgttcag aattggttaa
     4501 ttggttgtaa cactgacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4561 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gaatatgagt
     4621 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4681 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4741 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4801 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4861 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4921 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4981 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5041 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5101 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5161 gcgatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5221 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5281 cttccggctg gctggtttat tgctgataaa tccggagccg gtgagcgtgg ttctcgcggt
     5341 atcatcgcag cgctggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5401 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5461 attaagcatt ggtaagcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     5521 aaaatccctt aacgtgagtt acgcgcgcgt cgttccactg agcgtcagac cccgtagaaa
     5581 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5641 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5701 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5761 agttagccca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5821 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5881 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5941 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6001 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6061 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6121 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6181 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6241 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6301 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6361 cggaaggcga gagtagggaa ctgccaggca tcaaactaag cagaaggccc ctgacggatg
     6421 gcctttttgc gtttctacaa actctttctg tgttgtaaaa cgacggccag tcttaagctc
     6481 gggccccctg ggcggttctg ataacgagta atcgttaatc cgcaaataac gtaaaaaccc
     6541 gcttcggcgg gtttttttat ggggggagtt tagggaaaga gcatttgtca gaatatttaa
     6601 gggcgcctgt cactttgctt gatatatgag aattatttaa ccttataaat gagaaaaaag
     6661 caacgcactt taaataagat acgttgcttt ttcgattgat gaacacctat aattaaacta
     6721 ttcatctatt atttatgatt ttttgtatat acaatatttc tagtttgtta aagagaatta
     6781 agaaaataaa tctcgaaaat aataaaggga aaatcagttt ttgatatcaa aattatacat
     6841 gtcaacgata atacaaaata taatacaaac tataagatgt tatcagtatt tattatgcat
     6901 ttagaataaa ttttgtgtcg cccttattcg actcactata gaagttccta ttctctagaa
     6961 agtataggaa cttcacttca ttttatttaa atttgcggcc gctatgtctg ataaaaggat
     7021 gtaacatagg caagctgctc gtgagtgttg agtacgaacc ttagatccaa atcacccgca
     7081 cccacggata tacttgcttg aatatacagt agt
//
